Total Estrogen Concentrations Research Articles

Spatiotemporal variations and risk assessment of estrogens in the water of the southern Bohai Sea: A comprehensive investigation spanning three years

The ubiquitous and adverse effects of estrogens have aroused global concerns. Natural and synthetic estrogens in 255 water samples from the southern Bohai Sea were analyzed over three years. Total estrogen concentrations were 11.0–268 ng/L in river water and 1.98–99.7 ng/L in seawater, with bisphenol A (BPA) and 17α-ethynylestradiol (EE2) being the predominant estrogens, respectively. Estrogen showed the highest concentrations in summer 2018, followed by spring 2021 and spring 2019, which was consistent with the higher estrogen flux from rivers during summer. Higher estrogen concentrations in 2021 than in 2019 were driven by the higher level of BPA, an additive used in personal protective equipment. Estrogen exhibited higher concentrations in the southern coast of the Yellow River Delta and the northeastern coast of Laizhou bay due to the riverine input and aquaculture. Estrogens could disturb the normal endocrine activities of organisms and edict high ecological risks (90th simulated RQT > 1.0) to aquatic organisms, especially to fish. EE2 was the main contributor of estrogenic potency and ecological risk, which requires special concern. This is the first comprehensive study of estrogen spatiotemporal variations and risks in the Bohai Sea, providing insights into the environmental behavior of estrogens in coastal regions.

Socioeconomic and seasonal effects on spatiotemporal trends in estrogen occurrence and ecological risk within a river across low-urbanized and high-husbandry landscapes

Estrogen pollution is a persistent issue in rivers. This study investigated the occurrence, spatiotemporal variation mechanisms, sources, and ecological risks of estrone (E1), 17β-estradiol (E2), estriol (E3), 17α-ethinylestradiol (EE2), diethylstilbestrol (DES), and bisphenol-A (BPA) in the waters of the Zijiang River, a tributary of the middle Yangtze River. The results revealed elevated detection frequencies and estrogen concentrations in the dry season compared to the wet season, mainly due to the precipitation dilution effect. Total estrogen concentration ranged from 21.2 to 97.5 ng/L in the dry season, which was significantly correlated to spatial distributions of animal husbandry and population. Among the estrogens studied in the river, E2, BPA, and EE2 were predominant. The collective sources of E1, E2, E3, and EE2 were traced back to human and husbandry excrement, whereas BPA emitted from daily life products, contributing to 55.5% and 42.7% of the total estrogen concentration, respectively. Particularly, the average and median E1, E2, and EE2 concentrations in the river exceeded the environmental quality standards of the European Union. The total estrogenic activity dominated by EE2 exceeded the 1 ng E2/L threshold, with levels exceeding 10 ng E2/L during the dry season. The risk quotients exhibited a high ecological risk of E1 and EE2 to fish and a moderate to high ecological risk of E1 to crustaceans, EE2 to mollusks, and E2 to fish. Therefore, E1, E2, and EE2 pollution of the river may lead to both high estrogenic potency and moderate or high ecological risk; thus, they should be considered priority pollutants in the river. These results yield valuable insights into the spatiotemporal change mechanisms, sources, and ecological risks of estrogens in river water of low-urbanization and rural watersheds.

Investigation of free and conjugated estrogen fate and emission coefficients in three duck farms.

Concentration animal feeding operation (CAFO) is an important source of environmental estrogen. However, to the best of our knowledge, the data on estrogen discharge during duck breeding and growth is insufficient. This study used liquid chromatography with tandem mass spectrometry (LC/MS/MS) to analyze the free and conjugated estrogen concentrations in the surface water, outlet water, groundwater, and duck manure/soil mixture at three duck farms in Taiwan. Natural estrogen species included estrone (E1), 17β-estradiol (E2), estriol (E3), estrone-3-sulfate (E1-3S), 17β-estradiol-3-sulfate (E2-3S), estrone-3-glucuronide (E1-3G), and 17β-estradiol-3-glucuronide (E2-3G), whereas synthetic estrogen included 17α-ethynylestradiol (EE2) and diethylstilbestrol (DES). This study showed that the total estrogen concentrations in the surface water and groundwater were 15.4 and 4.5ng/L, respectively, which constituted 56% and 58%, respectively, conjugated estrogen. From the pond to the outlet water, the total estrogen concentration decreased by 3.9ng/L (23% loss) in the duck farms. However, the estrogenic potency was slightly reduced from 0.91 to 0.88 E2 equivalent/L, showing a negligible decrease. From the pond to the outlet water, the field results showed that converting the conjugated estrogen into free estrogen in the duck farm-released water increased their environmental hazard. Primarily E1, with an average concentration of 0.9 ± 1.6ng/g, was present in the duck manure. The estrogen excreted by the ducks in the pond (from surface water to outlet water) was estimated to be 0.18kg/million head-year. Although the estrogen concentration in the duck farms was low, the environmental impact of CAFO should not be neglected.

Endocrine profiling of reproductive status and evidence of pseudopregnancy in the Pacific walrus (Odobenus rosmarus divergens).

Endocrine profiling is an increasingly utilized tool for detecting pregnancies in wild populations of mammals. Given the difficulty in calculating reproductive rates of Pacific walruses (Odobenus rosmarus divergens) the use of endocrine techniques for determining pregnancy rates could be particularly useful for management of the population. The goals of this study were to 1) determine if progesterone and total estrogen concentrations in ovarian tissues of female walruses could be used to determine reproductive state and 2) determine if walruses undergo a functional postpartum estrus, as is seen in other pinnipeds. Ovaries were collected from female walruses (n = 13) hunted in subsistence hunts by Alaska Native communities. Females were categorized as postpartum, full-term pregnant, pregnant diapause or unbred. Total estrogen concentrations were greatest in unbred (n = 2) and pregnant (n = 2) females. Progesterone concentrations were also nominally larger in unbred (n = 2) than pregnant (n = 2) and postpartum (n = 9) animals. Small samples sizes precluded the use of statistical comparisons among groups. Corpora lutea tissue samples in this study did not reflect the presence of a postpartum estrus in the month of May as postpartum females yielded lower total estrogen concentrations than unbred or pregnant animals. Both unbred animals were in a state of pseudopregnancy, which has not been physiologically described for this species before. The progesterone profiles in late (59 ng/g) and early (140 ng/g) pregnancy were lower than expected and fell within the range of the postpartum females (36–210 ng/g), suggesting low production of the hormone by the corpus luteum during these phases of pregnancy. Profiling reproductive hormones in free-ranging walruses demonstrates that an endocrine approach may be a valuable tool for determining reproductive status of females, however increased sample sizes and time of year must be considered to accurately separate pregnant versus pseudopregnant individuals.

Molecularly Imprinted Polymers and Magnetic Molecularly Imprinted Polymers for Selective Determination of Estrogens in Water by ESI-MS/FAPA-MS.

Qualitative and quantitative analysis of estrogens content in natural water is a difficult task. An important problem in the analysis of hormones in water is the quantitative determination of their individual species. Low detection limits and instability of estrogen derivatives are the main challenges. Magnetic molecularly imprinted polymers (mag-MIPs) in combination with Flowing Atmospheric-Pressure Afterglow Mass Spectrometry (FAPA-MS) were successfully used for analysis of estrogen hormones in water samples. The aim of the study was to obtain mag-MIPs selective to estrone (E1) and β-estradiol (E2) for solid phase extraction and pre-concentration of estrogens. Due to their superior analyte binding properties at low concentrations (0.03 g in 1 g of polymer structure) and possibility of magnetic separation, mag-MIPs were proven to be very convenient and efficient adsorbent materials. In addition, MS analyses were performed using two ionization sources: ESI- and FAPA-MS. For both estrogens, LOD was significantly lower for FAPA-MS analysis (0.135 μg L−1 for E1 and E2) than for ESI-MS analysis (27 μg L−1 for E1 and 13.6 μg L−1 for E2). The total estrogen concentration in the environmental water sample was determined as: cE1 = 0.271 μg L−1 and cE2 = 0.275 μg L−1.

Estrogen Metabolism in Postmenopausal Women Exposed In Utero to Diethylstilbestrol.

Background: Prenatal diethylstilbestrol (DES) exposure is associated with adverse reproductive outcomes and cancer of the breast and vagina/cervix in adult women. DES effects on estrogen metabolism have been hypothesized, but reproductive hormone concentrations and metabolic pathways have not been comprehensively described.Methods: Blood samples were provided by 60 postmenopausal women (40 exposed and 20 unexposed) who were participants in the NCI Combined DES Cohort Study, had never used hormone supplements or been diagnosed with cancer, had responded to the most recent cohort study questionnaire, and lived within driving distance of Boston University Medical School (Boston, MA). Parent estrogens and their metabolites were measured by high-performance liquid chromatography-tandem mass spectrometry. Age-adjusted percent changes in geometric means and associated 95% confidence intervals (CIs) between the exposed and unexposed were calculated.Results: Concentrations of total estrogens (15.3%; CI, -4.1-38.5) and parent estrogens (27.1%; CI, -8.2-76.1) were slightly higher in the DES-exposed than unexposed. Ratios of path2:parent estrogens (-36.5%; CI, -53.0 to -14.3) and path2:path16 (-28.8%; CI, -47.3-3.7) were lower in the DES exposed. These associations persisted with adjustment for total estrogen, years since menopause, body mass index, parity, and recent alcohol intake.Conclusions: These preliminary data suggest that postmenopausal women who were prenatally DES exposed may have relatively less 2 than 16 pathway estrogen metabolism compared with unexposed women.Impact: Lower 2 pathway metabolism has been associated with increased postmenopausal breast cancer risk and could potentially offer a partial explanation for the modest increased risk observed for prenatally DES-exposed women. Cancer Epidemiol Biomarkers Prev; 27(10); 1208-13. ©2018 AACR.

Impact of preimplantational oral low-dose estradiol-17β exposure on the endometrium: The role of miRNA.

Porcine conceptuses synthesize estrogens between Day 11 and 12 as signal for maternal recognition of pregnancy. A preimplantational estrogen exposure to pregnant gilts has been associated with embryonic losses and changes in endometrial mRNA expression. MicroRNAs (miRNAs) play a key role in the mRNA regulation by modulating the expression. Effects of estrogens on endometrial miRNAs have not been investigated in this context so far. Thus, we studied the endometrial expression profile of miRNAs in the pig at gestational Day 10 after daily estradiol-17β (E2) application starting at fertilization using either 0, 0.05 (ADI-acceptable daily intake), 10 (NOEL-no-observed-effect level) and 1,000 (high dose) µg E2/kg body weight/day, respectively. In endometrial homogenates, E2 (p < 0.001) and total estrogen concentrations (p < 0.001) were significantly increased, namely 28- and 160-fold, respectively, in the high dose group as compared to the control. Additionally, total estrogens were sixfold elevated in the NOEL group. Interestingly, high-throughput sequencing of small non-coding RNA libraries did not indicate any differentially expressed miRNAs between the treatment groups and the control group. The expression of 12 potential E2 target miRNAs investigated by RT-qPCR were equally unaffected. Thus, preimplantational E2 exposure resulted in significantly higher endometrial estrogen concentrations, but did not perturb the expression profile of endometrial miRNAs.

Sex hormone-binding globulin regulation of androgen bioactivity in vivo: validation of the free hormone hypothesis.

Sex hormone-binding globulin (SHBG) is the high-affinity binding protein for androgens and estrogens. According to the free hormone hypothesis, SHBG modulates the bioactivity of sex steroids by limiting their diffusion into target tissues. Still, the in vivo physiological role of circulating SHBG remains unclear, especially since mice and rats lack circulating SHBG post-natally. To test the free hormone hypothesis in vivo, we examined total and free sex steroid concentrations and bioactivity on target organs in mice expressing a human SHBG transgene. SHBG increased total androgen and estrogen concentrations via hypothalamic-pituitary feedback regulation and prolonged ligand half-life. Despite markedly raised total sex steroid concentrations, free testosterone was unaffected while sex steroid bioactivity on male and female reproductive organs was attenuated. This occurred via a ligand-dependent, genotype-independent mechanism according to in vitro seminal vesicle organ cultures. These results provide compelling support for the determination of free or bioavailable sex steroid concentrations in medicine, and clarify important comparative differences between translational mouse models and human endocrinology.

Parturition induction in ewes by a progesterone receptor blocker, aglepristone, and subsequent neonatal survival: Preliminary results

The clinical effects of aglepristone treatment to induce parturition in ewes and their newborns were reported. Three experimental groups were defined: group AG5 (n = 5), group AG10 (n = 5), and group CG (n = 5) in which ewes were injected twice with 5, 10 mg/kg of aglepristone, and saline solution of ewes, respectively. Different parameters associated with parturition in ewes and their newborns were investigated. Serum progesterone, oxytocin, and free and conjugated total estrogens were measured after treatments until parturition. No statistical difference was found from first aglepristone administration to onset of lambing between AG5 and AG10 (23.90 ± 6.20, 40.00 ± 6.71 hours). Parturition induction in two groups shortened the gestational length significantly compared with the control group (P = 0.003). Dystocia was observed in two ewes in group AG10. The placental weight showed statistically significant difference only between the AG10 and CG (P = 0.039), but no difference was observed in the placental expulsion period between the groups. Decrease in food consumption 24 to 36 hours after parturition in all ewes and skin necrosis in an ewe in group AG5 were observed. Progesterone concentration was significantly lower in AG5 than that in ewes in group AG10 and CG (P < 0.05). No difference was observed in concentrations of free total estrogens and oxytocin between groups. The body temperature of lambs was significantly different between AG10 and CG groups both right after (P = 0.011) and 12 hours after parturition (P = 0.014). The lambs in CG had the highest mean birth weight (4.29 ± 0.28 kg), which was significantly different from the induced groups. No significant difference of blood pH and blood gases values between groups was identified both at birth and 12 hours after parturition for lambs. Significant differences could clearly be observed in total protein and blood urea nitrogen and total protein findings 12 hours after parturition (P < 0.05), whereas no difference was found in blood glucose, albumin, inorganic phosphor, triglyceride, or total cholesterol parameters. The results of this study show that the administration of aglepristone to induce parturition can precisely control lambing time without any side effects in either mothers or lambs.

Occurrence, distribution and risk assessment of estrogens in surface water, suspended particulate matter, and sediments of the Yangtze Estuary

The occurrence and distribution of six selected estrogen compounds were investigated in samples of surface water, suspended particulate matter (SPM), and sediment in the Yangtze Estuary and its coastal areas over four seasons. With the exception of 17α-ethinylestradiol (EE2), all estrogens were detected at least once in all three phases with bisphenol A (BPA) and estriol (E3) as the dominant estrogens in all phases. EE2 was not detected in any surface water samples. In addition, the highest total estrogen concentrations were found in January in all phases, which could be due to the low flow conditions and temperature during this season. A significant positive correlation was found between total estrogen concentrations and organic carbon (OC) contents, both in the water phase and solid phase (i.e. SPM and sediment), indicating the vital role played by OC. Based on a yeast estrogen screen (YES) bioassay, the higher estrogenic risk was found in the SPM and sediment phase when compared to the water phase. These results were confirmed by a risk assessment which revealed that the Yangtze Estuary was displayed a low to high risk over the seasons for all selected estrogens.

Comparison of endocrine changes, timing of ovulations, ovarian follicular growth, and efficacy associated with Estradoublesynch and Heatsynch protocols in Murrah buffalo cows (Bubalus bubalis)

Poor estrus expression and the difficulty encountered in predicting the time of ovulation compromise the reproductive efficiency of Murrah buffalo cows. Synchronization of ovulation and timed artificial insemination are able to precisely control the time of ovulation and thus avoid the need for estrus detection. Recently, the Estradoublesynch protocol (administration of a PGF2α injection 2 days before Heatsynch protocol; GnRH 0, PGF2α 7, estradiol benzoate [EB] 8) was developed that precisely synchronized ovulation twice, i.e., after GnRH and EB injections and resulted in satisfactory pregnancy rates in Murrah buffaloes. The present study was conducted on 104 cycling and 31 anestrus buffaloes to compare (1) the endocrine changes, timing of ovulations, ovarian follicular growth, and efficacy of Estradoublesynch and Heatsynch protocols in cycling and (2) the efficacy of Estradoublesynch and Heatsynch protocols for the improvement of fertility in cycling and anestrus Murrah buffalo cows. Ovulation was confirmed after all GnRH and EB treatments by ultrasonographic examination at 2-hour intervals. Plasma progesterone and total estrogen concentrations were determined in blood samples collected at daily intervals, beginning 2 days before the onset of protocols until the day of second ovulation detection. Ovulatory follicle size was measured by ultrasonography at six time points (first PGF2α administration of Estradoublesynch protocol every 2 days before the onset of Heatsynch protocol, GnRH administration of both protocols, 2 hours before ovulation detection after GnRH administration of both protocols, second PGF2α injection of Estradoublesynch protocol, PGF2α injection of Heatsynch protocol, EB injection of both protocols and, 2 hours before ovulation detection after EB administration of both protocols). Plasma LH, total estrogen, and progesterone concentrations were determined in blood samples collected at 30-minute intervals for 8 hours, beginning GnRH and EB injections, and thereafter at 2-hour intervals until 2 hours after the detection of ovulation. The first ovulatory rate was significantly higher (P < 0.05) in the Estradoublesynch protocol (84.6%) than that in the Heatsynch protocol (36.4%). The first LH peak concentration (74.6 ± 10.4 ng/mL) in the Estradoublesynch protocol was significantly higher (P < 0.05) than that of the Heatsynch protocol (55.3 ± 7.4 ng/mL). In Estradoublesynch protocol, the total estrogen concentration gradually increased from the day of GnRH administration coinciding with LH peak, and then gradually declined to the basal level until the time of ovulation detection. However, in Heatsynch protocol, the gradual increase in total estrogen concentration after GnRH administration was observed only in those buffalo cows, which responded to treatment with ovulation. In both Estradoublesynch and Heatsynch protocols, ovulatory follicle size increased by treatment with GnRH and EB until the detection of ovulation. The pregnancy rate after the Estradoublesynch protocol (60.0%) was significantly higher (P < 0.05) than that achieved after the Heatsynch protocol (32.5%). Satisfactory success rate using the Estradoublesynch protocol was attributed to the higher release of LH after treatment with GnRH, leading to ovulation in most of the animals and hence creating the optimum follicular size at EB injection for ovulation and pregnancy to occur.

A Bayesian network model for assessing natural estrogen fate and transport in a swine waste lagoon.

Commercial swine waste lagoons are regarded as a major reservoir of natural estrogens, which have the potential to produce adverse physiological effects on exposed aquatic organisms and wildlife. However, there remains limited understanding of the complex mechanisms of physical, chemical, and biological processes that govern the fate and transport of natural estrogens within an anaerobic swine lagoon. To improve lagoon management and ultimately help control the offsite transport of these compounds from swine operations, a probabilistic Bayesian network model was developed to assess natural estrogen fate and budget and then compared against data collected from a commercial swine field site. In general, the model was able to describe the estrogen fate and budget in both the slurry and sludge stores within the swine lagoon. Sensitivity analysis within the model demonstrated that the estrogen input loading from the associated barn facility was the most important factor in controlling estrogen concentrations within the lagoon slurry storage, whereas the settling rate was the most significant factor in the lagoon sludge storage. The degradation reactions were shown to be minor in both stores based on prediction of average total estrogen concentrations. Management scenario evaluations demonstrated that the best possible management options to reduce estrogen levels in the lagoon are either to adjust the estrogen input loading from swine barn facilities or to effectively enhance estrogen bonding with suspended solids through the use of organic polymers or inorganic coagulants.

Occurrence of free estrogens, conjugated estrogens, and bisphenol A in fresh livestock excreta and their removal by composting in North China.

An efficient pretreatment and analytical method was developed to investigate the occurrence and fate of four free estrogens (estrone (E1), 17β-estradiol (17β-E2), estriol (E3), and 17α-ethinylestradiol (EE2)), four conjugated estrogens (estrone-3-sulfate sodium salt (E1-3S), 17β-estradiol-3-sulfate sodium salt (E2-3S), estrone-3-glucuronide sodium salt (E1-3G), and 17β-estradiol-3-glucuronide sodium salt (E2-3G)), and bisphenol A (BPA) in three livestock farms raising beef cattle, cows, sheep, swine, and chickens in Qi County, which is located in North China. The results demonstrated that one cow and one beef cattle excreted 956.25-1,270.41 and 244.38-319.99 μg/day of total (free and conjugated) estrogen, respectively, primarily through feces (greater than 91%), while swine excreted 260.09-289.99 μg/day of estrogens, primarily through urine (98-99%). The total estrogen excreted in sheep and broiler chicken feces was calculated to be 21.64-28.67 and 4.62-5.40 μg/day, respectively. It was determined that conjugated estrogens contributed to 21.1-21.9% of the total estrogen excreted in cow feces and more than 98% of the total estrogen excreted in swine urine. After composting, the concentration of total estrogen decreased by 18.7-59.6%; however, increased levels of BPA were measured. In treated compost samples, estrogens were detected at concentrations up to 74.0 ng/g, which indicates a potential risk of estrogens entering the surrounding environment.

Endocrine changes, timing of ovulation, ovarian follicular growth and efficacy of a novel protocol (Estradoublesynch) for synchronization of ovulation and timed artificial insemination in Murrah buffaloes (Bubalus bubalis)

Experiments were conducted to investigate (1) ovarian follicular growth, timing of ovulation and associated endocrine changes (progesterone, estrogen, and LH) in cycling, and (2) efficacy in terms of pregnancy rate in cycling and anestrus Murrah buffaloes subjected to the Estradoublesynch protocol (prostaglandin F2α [PGF2α] 0, GnRH 2, PGF2α 9, estradiol benzoate, EB 10). Twelve cycling buffaloes were subjected to the Estradoublesynch protocol and observed for ovulation, follicle size, and endocrine changes after EB treatment. Ovulation occurred in 12 of 12 buffaloes (100%) at 48.5 ± 1.6 hours (range, 38.0–56.0) after EB treatment. Plasma LH, total estrogen, and progesterone concentrations were determined in intensive blood samples collected after EB treatment. Peak LH concentration of 34.2 ± 7.7 ng/mL (range, 17.8–178.5) occurred at 18.3 ± 0.8 hours (range, 14.0–22.0) after EB treatment. Peak total estrogen of 50.8 ± 6.9 pg/mL (range, 32.3–82.7) occurred 5.7 ± 1.0 hours (range, 2.0–14.0) after EB treatment. Follicle size was gradually increased from second PGF2α injection (9.7 ± 0.3 mm; range, 8.0–12.0) until ovulation was detected (12.9 ± 0.4 mm; range, 11.0–15.0). Fourteen cycling and 11 anestrus buffaloes were subjected to the Estradoublesynch protocol, with timed artificial insemination (TAI) 48 and 60 hours after EB treatment, and 58 cycling buffaloes were inseminated after spontaneous estrus (control group). Pregnancy rates were 62% for TAI of cycling buffaloes, 64% for anestrus buffaloes, and 34.5% for control group. Our observations demonstrated that the Estradoublesynch protocol followed by TAI satisfactory enhanced pregnancy rates in both cycling and anestrus buffaloes.

Effects of a protracted induction of parturition on the incidence of retained placenta and assessment of uterine artery blood flow as a measure of placental maturation in cattle

The objectives of the present study were to compare the effects of a protracted and a conventional induction of parturition on the incidence of retained placenta, and to evaluate the suitability of transrectal Doppler sonography of the uterine arteries as a noninvasive method for the assessment of placental maturation. Protracted induction of labor (PIP) was precipitated in 13 cows by the administration of 1.3 mg dexamethasone im twice daily between Days 268 and 273 of gestation, and 40 mg dexamethasone im on Day 274 of gestation. For conventional induction of labor (SIP), 10 cows received 40 mg dexamethasone on Day 274 of gestation. A third group was not treated and served as control (SPON; N = 11). Blood flow volume (BFV) and resistance index in the uterine arteries were measured with Doppler sonography once a day from Day 268 of gestation until labor. After each ultrasonographic examination, blood samples for determination of steroid hormones were taken. Incidence of retained placenta was lower (P < 0.05) in group SPON (9%) compared with groups PIP (54%) and SIP (70%). In the last 7 days before parturition uterine BFV and resistance index did not change (P > 0.05) and did not differ between groups SPON, PIP, and SIP (P > 0.05). Resistance index was higher (P < 0.001) in cows with retained placenta compared with cows with released placenta, and BFV did not differ (P > 0.05) between them. Total estrogen concentrations increased by 283% (P < 0.001) in group PIP and by 60% (P < 0.05) in group SPON between Days −7 and −1 before parturition. They stayed constant (P > 0.05) until Day −2 in group SIP, but increased (P < 0.05) after the high dosage of dexamethasone within 1 day by 140%. Total estrogen levels were higher (P < 0.05) in cows with released placenta than in cows with retained placenta. In conclusion, a protracted compared with a short induction of labor results in higher estrogen levels before term, but does not affect incidence of placental retention. Neither alterations in placental maturation nor changes in steroid hormones influenced uterine blood supply. Therefore, Doppler sonography of uterine arteries is unsuitable to investigate the process of placental maturation induced by glucocorticoids in cows. Nevertheless, disturbances in the placental maturation process in cows with retained fetal membranes after parturition can be detected before parturition by a higher uterine blood flow resistance in the uterine arteries.

The endocrine changes, the timing of ovulation and the efficacy of the Doublesynch protocol in the Murrah buffalo (Bubalus bubalis)

Experiments were conducted to investigate (a) the timing of ovulation and the associated endocrine changes (progesterone, estrogen and LH) during estrous cycle and (b) the efficacy, with respect to the pregnancy rate, in cycling and anestrus in Murrah buffaloes subjected to the Doublesynch protocol during the low breeding season. In experiment 1, 10 cycling buffaloes were administered PGF2α on day 0 (without regard to the estrous cycle stage), GnRH on day 2, a second PGF2α injection on day 9, and a second GnRH injection on day 11. Transrectal palpation was performed at 2-h intervals after the first and second GnRH treatments until ovulation was detected or for upto 96h. The plasma progesterone and total estrogen concentrations were determined in blood samples collected at daily intervals starting 2days before the onset of the protocol and continued until the day of the second detected ovulation. The plasma LH and total estrogen concentrations were measured in blood samples collected at 30-min intervals for 8h following the first and second GnRH injections and thereafter at 2-h intervals until 2h after the detection of ovulation. Ovulation occurred in 9/10 buffaloes (90%) at 22.2±1.2h (mean±S.E.M.; range 18.0–26.0h) and 10/10 buffaloes (100%) at 23.2±1.0h (mean±S.E.M.; range 20.0–28.0h) after the first and second GnRH treatments, respectively. The peak LH concentrations of 99.8±28.5ng/ml (range 37.8–320.0ng/ml) and 62.3±11.9ng/ml (range 20.9–143.9ng/ml) occurred 2.1±0.3h (range 1.0–3.5h) and 2.3±0.3h (range 0.5–3.0h) after the first and second GnRH treatments, respectively. The total estrogen concentration gradually increased from the day of both the first and second PGF2α administrations until the LH peak (with great variability) and then gradually declined to the basal level, which was reached at the time ovulation was detected. In experiment 2, 10 cycling and 11 non-lactating anestrus buffaloes were subjected to the Doublesynch protocol with timed artificial insemination (TAI) 16 and 24h after the second GnRH treatment, and 55 cycling buffaloes were inseminated after spontaneous estrus was detected (control group). The pregnancy rates were 60% using TAI on cycling buffaloes (experiments 1 and 2), 55% for anestrus buffaloes (experiment 2), and 27.3% for cycling buffaloes inseminated following spontaneous estrus. The overall pregnancy success rates after the Doublesynch protocol in both cycling and anestrus buffaloes increased by 30.8% compared to spontaneous estrus (58.1% vs. 27.3%).In conclusion, the Doublesynch protocol effectively synchronized ovulation twice (after the first and second GnRH treatments) irrespective of the stage of estrous cycle in Murrah buffaloes. The study also demonstrated that the Doublesynch protocol followed by TAI significantly (P<0.005) enhanced the pregnancy rate in cycling and anestrus buffaloes in comparison to untreated controls during the low breeding season.

Dissolved Organic Carbon and Estrogen Transport in Surface Runoff From Agricultural Land Receiving Poultry Litter1

Dutta, Sudarshan, Shreeram Inamdar, Jerry Tso, Diana S. Aga, and J. Tom Sims, 2012. Dissolved Organic Carbon and Estrogen Transport in Surface Runoff from Agricultural Land Receiving Poultry Litter. Journal of the American Water Resources Association (JAWRA) 48(3): 558‐569. DOI: 10.1111/j.1752‐1688.2011.00634.xAbstract: Dissolved organic carbon (DOC) provides a reactive substrate for the transport of organic contaminants with runoff. Very few studies have investigated the export of DOC from agricultural land, especially those receiving manure applications. We investigated exports of DOC in surface runoff from agricultural fields receiving various treatments of poultry litter (raw vs. pelletized). In addition, we also investigated how estrogens in runoff were associated with DOC. Different forms of estrogens studied were: estrone, 17β‐estradiol, estriol, and their conjugates. Experimental agricultural plots were 12 m × 5 m long and had reduced tillage and no‐till management practices. The aromatic content of DOC was characterized using specific ultraviolet absorbance (SUVA). Flow‐weighted concentrations of DOC and SUVA in surface runoff from plots with poultry litter were significantly (p ≤ 0.10) greater than the control (no litter) plots. Compared to pelletized poultry litter, reduced‐tillage plots with raw litter yielded higher DOC concentrations and SUVA values. No significant differences (p ≥ 0.10) in DOC and SUVA were observed between litter treatments for plots with no‐till. Total estrogen concentrations (including all forms) were positively and significantly (p ≤ 0.10) correlated with DOC. These results can help select and guide agricultural management practices that can reduce the exports of DOC and associated contaminant from agricultural land receiving manure applications.

The intravaginal application of misoprostol improves induction of abortion with aglepristone

The aim of the present study was to compare the clinical and endocrinological effects of four different treatments for the induction of abortion in bitches. For this purpose, 28 pregnant bitches between days 25 and 35 of gestation, were randomly assigned to four groups. In group I (n = 7), only aglepristone (AGL, 10mg/kg bw, two injections 24 h apart, s.c.) was administered. In group II (n = 7), AGL (as in group I), cabergolin (CAB, 5 μg/kg, daily p.o., until completion of abortion) and misoprostol (MIS, 200 μg for bitches with ≤ 20 kg bw, 400 μg for bitches with > 20 kg bw, daily intravaginally, until completion of abortion) were administered. In group III (n = 7), AGL (as in group I) and MIS (as in group II) were administered. In group IV (n = 7) AGL, (as in group I) and cloprostenol (CLO, 1μg/kg bw, s.c., two injections 24 h apart with the AGL injections) were combined. In all groups, bitches were examined daily, clinically and ultrasonographically to monitor resorptions/abortions. To measure serum progesterone (P4) and total estrogen (TE) concentrations, blood samples were collected in all groups immediately after the first AGL administration and every other day until completion of abortion. No statistical differences were found between groups concerning the duration until completion of abortion following treatment (n.s.); however, in Group III, 6 d after the start of treatment all pregnancies were terminated whereas in Group I, II and IV, only 57.1% (4/7), 85.7 % (6/7) and 42.8 % (3/7) of pregnancies were terminated. In the latter groups, all pregnancies were terminated between days 8 and 10 after the start of treatment. In Group IV, P4 concentrations on days two and one before the beginning of abortion and the day the abortion started was significantly lower than in the other groups (P < 0.01). No statistical differences were found between groups for TE concentrations (P > 0.05). In Groups I, II and III, no severe side effects occurred. Severe vomiting after each treatment and until the end of abortion was observed in Group IV only. In conclusion, only when a combination of AGL and MIS was used abortion was completed within 6 d in all bitches whereas the additional use of CAB did not improve the treatment.

A Comparison of Recombinant Receptor-Reporter Gene Bioassays and a Total Estrogen Enzyme Linked Immunosorbent Assay for the Rapid Screening of Estrogenic Activity in Natural and Waste Waters

In this study, we compared the performance as screening tools of two yeast-based recombinant receptor-reporter gene bioassays with a commercial ELISA kit for measurement of total estrogens. For WWTP effluents there was a very good correlation between the measured total estrogen concentrations (ELISA) and estrogenic activity by the hERα bioassay (r(2)=0.93), but not for the medERα bioassay (r(2)=0.50). For freshwater samples, the correlations between bioassay response and ELISA ES measurements were very good (r(2)>0.95). There was no correlation between bioassay response and ELISA ES measurements for estuarine samples.

Passive Secondary Biological Treatment Systems Reduce Estrogens in Dairy Shed Effluent

Steroid estrogens are found at high concentrations in untreated dairy shed effluents. Reduction of estrogenic activity and steroid estrogen concentrations was assessed in two systems used to treat dairy shed effluents: the two-pond system and the advanced pond system. Both include anaerobic and aerobic treatment stages. Samples of effluent were collected from the systems and analyzed for free estrogens, conjugated estrogens and total estrogenic activity using E-Screen assay. Both systems showed increases of up to 8000% in aqueous free estrogens and estrogenic activity after anaerobic treatment, followed by decreases after aerobic treatment (36-83%). The complete systems decreased total steroid estrogen concentrations by 50-100% and estrogen activity by 62-100%, with little difference between systems. Removal rates were lower in winter for both systems. Final effluents from the advanced pond system contained total estrogens at <15-1400 ng/L and estrogenic activity at 3.2-43 ng/L. Final effluent from the two-pond system contained total estrogens at <15-300 ng/L and estrogenic activity at 3.3-25 ng/L. At times the final effluent EEQs exceeded guideline values for protection of freshwater fish and suggest further treatment may be required.